Bacterioria test material

ABSTRACT

An improved bacteriuria test material is made up of an admixture of sulfanilamide and N-(1-Naphthyl) ethylene-diamine dihydrochloride. In one embodiment a solution of these materials is employed to impregnate a paper strip or a cotton swab and then permitted to dry. In use when the resulting test material, such as the aforementioned impregnnated paper or cotton swab, is brought into contact or wetted with a test specimen containing nitrite, indicating the presence of bacteria in the urine or the test specimen, the paper or swab undergoes a fairly rapid color change to a rose or red color.

United States Patent [191 Lagomarsino [451 Feb. 27, 1973 BACTERIORIATEST MATERIAL 75] Inventor: John A. Lagomarsino, Wantagh,

[73] Assignee: Bio-Derivatives Corporation, Deer Park,N.Y.

[22] Filed: Jan.4, 1971 [21] Appl. No.: 103,695

[52] U.S. Cl ..l95/99, 195/l03.5 R, 195/127 [51] Int. Cl. ..C12k 1/10[58] Field of Search....195/103.5 R, 100; 23/253 TP; 252/408 [56]References Cited UNITED STATES PATENTS 3,415,717 12/1968 Avakian..l95/l00 3,547,780 12/1970 Finnerty ..195/l03.5 R

3,232,710 2/1966 Rieckmann et al. ..195/l03.5 R

3,634,198 1/l972 Truhan ..195/l03.5 R

Primary Examiner-Lionel M. Shapiro Assistant Examiner-Robert J. WardenAttorney-Robert S. Dunham, P. E. Henninger, Lester W. Clark, Gerald W.Griffin, Thomas F. Moran, Howard J. Churchill, R. Bradlee Boa],Christopher C. Dunham, Robert Scobey and Henry T. Burke [5 7 ABSTRACT Animproved, bacteriuria test material is made up of an admixture ofsulfanilamide and N-(l-Naphthyl) ethylene-diamine dihydrochloride. Inone embodiment a solution of these materials is employed to imprcgnate apaper strip or a cotton swab and then permitted to dry. In use when theresulting test material, such as the aforementioned impregnnated paperor cotton swab, is brought into contact or wetted with a test specimencontaining nitrite, indicating the presence of bacteria in the urine orthe test specimen, the paper or swab undergoes a fairly rapid colorchange to a rose or red color.

20 Claims, 5 Drawing Figures PATENTED 3,718,543

- INVENTOR.

VZ JOHN A. LAGOMARSINO ATTORNEY BACTERIORIA TEST MATERIAL This inventionrelates to test materials or reagents useful in determining bacteriuria.More particularly, Griess, see Ber. deutsch. chem. Gesellsch. i2,426-428 (1879), shows that certain bacteria could be detected in watersupplies due to the presence of nitrite by the production of a red colorupon addition of an acidic sulfanilic acid-alpha-naphthylamine reagent.U.S. Pat. No. 3,415,717 discloses a bacteriuria test paper embodying theGriess test and made up of paper impregnated with a sulfonamide, anN,N-dialkyl-lnaphthylamine and an acyclic hydroxy polycarboxylic acid.Such test papers, however, must be stored in a light-proof, air-tightbottle preferably in the presence of a desiccant. Moreover, such testmaterials are made up of three components, a sulfonamide, an appropriateamine and an acid.

It is an object of this invention to provide improved reagent materialuseful in determining the presence of bacteriuria.

It is another object of this invention to provide an improvedbacteriuria cotton test swab and an improved bacteriuria test paper andthe like.

How these and other objects of this invention are accomplished willbecome apparent in the light of the accompanying disclosure made withreference to the accompanying drawing wherein there are illustratedvarious embodiments of the practices of this invention, specifically,

Wherein FIG. 1 illustrates a test paper impregnated with the combinationof test ingredients in accordance with this invention;

Wherein FIG. 2 illustrates a protected test paper in accordance with thepractices of this invention;

Wherein FIG. 3 is a cross-sectional view taken along line 3-3 of FIG. 2;

Wherein FIG. 4 illustrates a test swab impregnated with the combinationof test ingredients in accordance with this invention; and

Wherein FIG. 5 illustrates a protected test swab in accordance with thisinvention.

An improved bacteriuria test reagent in accordance with this inventioncomprises a sulfanilamide and N-( l- Naphthyl)ethylenediaminedihydrochloride. In accordance with one embodiment a bacteriuria testpaper or bacteriuria cotton swab and the like is provided by forming asolution containing a sulfanilamide, such as the compound sulfanilamide,and N-( 1- Naphthyl)ethylenediamine dihydrochloride and impregnating asuitable substrate, such as paper or a cotton swab, with the solutionand then permitting the resulting impregnated substrate, such as thepaper or swab, to dry. When the resulting test paper or swab or even thesolution itself is brought into contact with a test specimen todetermine the presence of bacteriuria, a color change of the test paperor swab to a reddish color occurs, if the test is positive, i.e., ifthere is bacteriuria, since bacteriuria leads to the formation ofnitrites which upon contact with the aforesaid test materials bringsabout the above-mentioned color change.

Various solvents may be employed in the preparation of the impregnatedtest paper or test swabs in accordance with this invention. For example,a concentrated or saturated or substantially saturated acetone solutionof sulfanilamide is formed. Other suitable solvents might be employed inplace of or in admixture with acetone. There is also formed aconcentrated or saturated or substantially saturated N-( l-Naphthyl)ethylenediamine dihydrochloride in alcohol or other suitablesolvent. These solutions could then be combined and the resultingsolution comprising an admixture of sulfanilamide and N-( l-Naphthyl)ethylenediamine dihydrochloride could then be employed toimpregnate the test substrate materials, such as the test paper, cottonswab or the like.

If desired, a single solvent, a ketone, such as acetone, an alcohol,such as ethanol, an ether, such as ethyl ether, or a mixture ofsolvents, e.g., the aforementioned solvents, might be employed as thecommon solvent for the combination of the active test materials inaccordance with this invention. For example, an acetone and/or ethanolsolution of sulfanilamide and N-( l- Naphthyl)ethylenediaminedihydrochloride could be prepared and the resulting solution employed toimpregnate the test paper or cotton swab or other suitable substrate orbacteriuria test carrier. Solutions containing from about 2% by weightor less up to about 10 percent by weight or more of the aforementionedtest components, separately or together are usefully employed in thepractice of this invention for the preparation of the bacteriuria testreagent materials, particularly impregnated bacteriuria test papers andthe impregnated bacteriuria cotton test swabs.

Desirably, in the preparation of the bacteriuria test reagents inaccordance with this invention, the sulfanilamide component, e.g.,sulfanilamide and the other component N-(l-Naphthyl)ethylenediaminedihydrochloride, are present in a stoichiometric amount relative to eachother. For example, if desired, these materials are present in admixturein the bacteriuria test reagent on an equimolar basis. If desired,however, on a molar basis one component may be present in a greateramount than the other component. Further, by way of example, thesulfanilamide component may be present relative to the N-( l-Naphthyl)ethylenediamine dihydrochloride component in a weight or molarratio in the range 1:10 or from 10:1, respectively. A broad range of theproportions of the test components making up the bacteriuria reagentmaterials in accordance with this invention is possible due to theintense and noticeable color change brought about by the presence ofthese materials in contact with an aqueous nitrite-containing solution.The intensity of the change in color to a red color is indicative notonly of the presence of bacteriuria but may also be indicative of theextent of the bacteria infection.

In the practice of this-invention it is preferred that the components ofthe bacteriuria test reagent be incorporated or impregnated on a paperstrip or cotton swab. Desirably, the cotton swab is disposed or wrappedor fixed to the end of a suitable stick or carrier, e.g., in the natureof Q-Tips cotton swabs manufactured by Cheseborough-Ponds, Inc. NewYork, N. Y.

As indicated hereinabove, the impregnated bacteriuria test paper or swabis immersed in the test specimen to determine by an observable colorchange in the test paper or swab from a white or natural color,depending upon the nature of the substrate material, to a reddish colorindicative of the presence of nitrite and therefore a bacteria infectionor bacteriuria if the test specimen is a urine sample. If desired,however, the test reagents in accordance with this invention may beemployed in solution form. In this embodiment a solution of the testreagents is admixed with the test specimen and the color changeobserved. The advantages of this invention, however, are obtainable whenthe bacteriuria test materials of this invention are incorporated orimpregnated in dry form in a suitable carrier, such as a paper or cottonswab.

In accordance with yet another embodiment of the practices of thisinvention, instead of forming a solution containing the test reagents inaccordance with this invention, a finely divided dry admixture of thetest reagents may be formed and then coated onto or otherwise affixed tothe paper or cotton swab carrier. Instead of forming a dry, finelydivided admixture of these materials, the materials in finely dividedform may be separately applied to the carrier.

Various embodiments of the practices of this invention are illustratedin the accompanying drawing. Specifically, FIG. 1 illustrates a strip 10of test paper impregnated with a combination of test reagents, e.g.,sulfanilamide and N-( l-Naphthyl)ethylenediamine dihydrochloride inaccordance with this invention. FIGS. 2 and 3 illustrate a strip of testpaper, of the type illustrated in FIG. 1, wherein the test paper issandwiched between or protected on both sides by a removable orstrippable film or layer or coating 11 of substantiallymoisture-impermeable plastic material, such as a thin film ofpolyethylene or Mylar or polyvinyl chloride or the like. Film 11 ofplastic material serves to protect the underlying or sandwiched testpaper from exposure to moisture and the surrounding atmosphere and toprotect test paper 10 during storage and/or handling.

FIG. 4 illustrates a test swab in accordance with this invention whereinwad 12 of cotton impregnated with the test reagent in accordance withthis invention is gathered around and fixed to one end of a suitablecarrier or stick 14. FIG. 5 is illustrative of a special embodiment of atest swab in accordance with this invention as illustrated in FIG. 2. Inthe embodiment illustrated in FIG. 5 cotton wad 12 is protected in themanner similar to test paper as illustrated in FIGS. 2 and 3 byenclosing wad 12 within a removable, substantially moisture-impermeableplastic coating, film or envelope which is suitably affixed to holder orstick 14 as illustrated, e.g., by twistedly affixing film 15 to holder14.

The following example is illustrative of the practices of thisinvention. A substantially saturated acetone solution of sulfanilamideis formed. There is also formed a solution ofN-(l-Naphthyl)ethylenediamine dihydrochloride in ethanol. Thesesolutions are then combined. A suitable substrate material, such aspaper or a cotton swab, is then wetted with or introduced into thecombined solutions and dried. When employed in a bacteriuria testprocedure the impregnated paper or swab is brought into contact with orimmersed in the test specimen. In a positive test, i.e., an indicationof nitrite due to the presence of bacteria, the test paper or swab willchange from a white to dark red, light red, pink or purple color. Any ofthese color changes indicates a positive reaction and the presence ofbacteriuria. Upon the observation of a positive test or reaction thetest specimen is then sent to a diagnostic laboratory for bacteriacolony count and for identification of the bacteria. From theselaboratory tests a diagnosis can be rendered and appropriate therapyinitiated.

Other embodiments of the practices of this invention include acompressed admixture of the reagent materials, such as a sulfanilamide,e.g., sulfanilamide, and N- (l-Naphthyl)ethylenediamine dihydrochloride,the compressed admixture preferably being in the form of a tablet ordisc. The compressed admixture would desirably include an inert carrieror diluent, preferably white in color and water-insoluble, which wouldalso usefully function as a binder for the aforesaid reagent materials.Suitable such carrier materials or diluents would include the normallywater-insoluble inert salts, such as in tablet or disc form, would thenhave added thereto a drop of the urine specimen to be tested.

In accordance with another embodiment of this invention involving apaper impregnated with the special reagent admixture, paper, preferablycut into a desired shape, such as a strip or circle, is coated orimpregnated with the reagent admixture. In use, the paper is contactedwith the drop of urine specimen to be tested.

As will be apparent to those skilled in the art in the light of theforegoing disclosure, many modifications, alterations and substitutionsare possible in the practice of this invention without departing fromthe spirit or scope thereof.

I claim:

1. A composition useful as a bacteriuria test material consistingessentially of an admixture of a sulfanilamide and N-(l-Naphthyl)ethylenediamine dihydrochloride.

2. A composition in accordance with claim 1 wherein said admixture iscompressed into tablet or disc or wafer form.

3. A composition in accordance with claim 1 wherein said admixturecontains a major amount of a whitecolored, water-insoluble, inert,finely divided carrier or diluent, the resulting admixture beingcompressed into tablet form. v

4. A composition in accordance with claim 1 wherein said sulfanilamideis sulfanilamide.

5. A test material in accordance with claim 1 wherein said admixture isdisposed in an anhydrous carrier.

6. A test material in accordance with claim 5 wherein said carrier is analcohol.

7. A test material in accordance with claim 5 wherein said carrier is aketone.

8. A test material in accordance with claim 5 wherein said carrier is asolid carrier.

9. A test material in accordance with claim 5 wherein said carrier is apaper.

10. A test material in accordance with claim 9 wherein said paper isprotected by a strippable substantially moisture-impermeable film orcoating.

11. A test material in accordance with claim 5 wherein said carrier is acotton swab.

12. A test material in accordance with claim 11 wherein said cotton swabis affixed to a wooden or plastic holder.

13. A test material in accordance with claim 12 wherein said cotton swabis enclosed within a removable substantially moisture-impermeableenvelope or coating.

14. A test material in accordance with claim 5 wherein said carrier is asolid, porous or permeable water-insoluble carrier.

15. In a bacteriuria test wherein the specimen to be tested is broughtinto contact with a reagent material which undergoes a change in color,the improvement which comprises employing as said reagent a materialwhich consists essentially of a sulfanilamide and N-(l-Naphthyl)ethylenediamine dihydrochloride.

16. A method in accordance with claim wherein said reagent consistsessentially of sulfanilamide and N- (l -Naphthyl )ethylenediaminedihydrochloride disposed on a porous, water-insoluble carrier.

17. A method in accordance with claim 16 wherein said carrier is paper.

18. A method in accordance with claim 16 wherein said carrier is cottonswab.

19. A method in accordance with claim 18 wherein said sulfanilamide andsaid N-( l- Naphthyl)ethylenediamine dihydrochloride are present in therelative proportions in parts by weight or mols in the range 1:10 to10:1, respectively.

20. A bacteriuria test paper consisting essentially of paper impregnatedwith a sulfanilamide and N-( l- Naphthyl) ethylenediaminedihydrochloride wherein said sulfanilamide and said N-( 1-Naphthyl)ethylenediamine dihydrochloride are present in the relativeproportions in parts by weight or mols in the range 1:10 to 10:1,respectively.

2. A composition in accordance with claim 1 wherein said admixture iscompressed into tablet or disc or wafer form.
 3. A composition inaccordance with claim 1 wherein said admixture contains a major amountof a white-colored, water-insoluble, inert, finely divided carrier ordiluent, the resulting admixture being compressed into tablet form.
 4. Acomposition in accordance with claim 1 wherein said sulfanilamide issulfanilamide.
 5. A test material in accordance with claim 1 whereinsaid admixture is disposed in an anhydrous carrier.
 6. A test materialin accordance with claim 5 wherein said carrier is an alcohol.
 7. A testmaterial in accordance with claim 5 wherein said carrier is a ketone. 8.A test material in accordance with claim 5 wherein said carrier is asolid carrier.
 9. A test material in accordance with claim 5 whereinsaid carrier is a paper.
 10. A test material in accordance with claim 9wherein said paper is protected by a strippable substantiallymoisture-impermeable film or coating.
 11. A test material in accordancewith claim 5 wherein said carrier is a cotton swab.
 12. A test materialin accordance with claim 11 wherein said cotton swab is affixed to awooden or plastic holder.
 13. A test material in accordance with claim12 wherein said cotton swab is enclosed within a removable substantiallymoisture-impermeable envelope or coating.
 14. A test material inaccordance with claim 5 wherein said carrier is a solid, porous orpermeable water-insoluble carrier.
 15. In a bacteriuria test wherein thespecimen to be tested is brought into contact with a reagent materialwhich undergoes a change in color, the improvement which comprisesemploying as said reagent a material which consists essentially of asulfanilamide and N-(1-Naphthyl)ethylenediamine dihydrochloride.
 16. Amethod in accordance with claim 15 wherein said reagent consistsessentially of sulfanilamide and N-(1-Naphthyl)ethylenediaminedihydrochloride disposed on a porous, water-insoluble carrier.
 17. Amethod in accordance with claim 16 wherein said carrier is paper.
 18. Amethod in accordance with claim 16 wherein said carrier is cotton swab.19. A method in accordance with claim 18 wherein said sulfanilamide andsaid N-(1-Naphthyl)ethylenediamine dihydrochloride are present in therelative proportions in parts by weight or mols in the range 1:10 to10:1, respectively.
 20. A bacteriuria test paper consisting essentiallyof paper impregnated with a sulfanilamide and N-(1-Naphthyl)ethylenediamine dihydrochloride wherein said sulfanilamide and saidN-(1-Naphthyl)ethylenediamine dihydrochloride are present in therelative proportions in parts by weight or mols in the range 1:10 to10:1, respectively.